Temperature Effects on Expression Levels of hsp Genes in Eggs and Second-Stage Juveniles of Meloidogyne hapla Chitwood, 1949.

Meloidogyne hapla is one of the most important nematode pathogens. It is a sedentary, biotrophic parasite of plants that overwinters in the soil or in diseased roots. The development of M. hapla is temperature dependent. Numerous studies have been performed on the effect of temperature on the development of M. hapla, but only a few of them analyzed the heat shock protein (hsp) genes. The aim of the study was to perform expression profiling of eight hsp genes (Mh-hsp90, Mh-hsp1, Mh-hsp4, Mh-hsp6, Mh-hsp60, Mh-dnj19, Mh-hsp43, and Mh-hsp12.2) at two development stages of M. hapla, i.e., in eggs and second-stage juveniles (J2). The eggs and J2 were incubated under cold stress (5 °C), heat stress (35 °C, 40 °C), and non-stress (10 °C, 20 °C, and 30 °C) conditions. Expression profiling was performed by qPCR. It was demonstrated that only two genes, Mh-hsp60 and Mh-dnj19, have been upregulated by heat and cold stress at both development stages. Heat stress upregulated the expression of more hsp genes than cold stress did. The level of upregulation of most hsp genes was more marked in J2 than in eggs. The obtained results suggest that the Mh-hsp90 and Mh-hsp1 genes can be used as bioindicators of environmental impacts on nematodes of the Meloidogyne genus.

Organization of DNA Replication Origin Firing in Xenopus Egg Extracts: The Role of Intra-S Checkpoint.

During cell division, the duplication of the genome starts at multiple positions called replication origins. Origin firing requires the interaction of rate-limiting factors with potential origins during the S(ynthesis)-phase of the cell cycle. Origins fire as synchronous clusters which is proposed to be regulated by the intra-S checkpoint. By modelling the unchallenged, the checkpoint-inhibited and the checkpoint protein Chk1 over-expressed replication pattern of single DNA molecules from Xenopus sperm chromatin replicated in egg extracts, we demonstrate that the quantitative modelling of data requires: (1) a segmentation of the genome into regions of low and high probability of origin firing; (2) that regions with high probability of origin firing escape intra-S checkpoint regulation and (3) the variability of the rate of DNA synthesis close to replication forks is a necessary ingredient that should be taken in to account in order to describe the dynamic of replication origin firing. This model implies that the observed origin clustering emerges from the apparent synchrony of origin firing in regions with high probability of origin firing and challenge the assumption that the intra-S checkpoint is the main regulator of origin clustering.

Assessment of Embryonic Bioactivity through Changes in the Water Structure Using Near-Infrared Spectroscopy and Imaging.

We explored the influence of embryonic bioactivity on the water structure using near-infrared (NIR) spectroscopy and imaging. Four groups of Japanese medaka fish (Oryzias latipes) eggs were studied: (a) one group of eggs was activated by fertilization, and (b-d) three groups of eggs were not activated because embryogenesis was stopped or not started by (b) culturing under cold temperature, (c) instant freezing, or (d) lack of fertilization. The yolks of the activated eggs contained higher proportions of weakly hydrogen bonded water than those of nonactivated eggs. A possible factor responsible for the significant changes in the water structure was revealed to be a protein secondary structural change from an α-helix to a ß-sheet in the activated eggs. NIR images of the activated eggs successfully visualized the water structural variation in the yolk with a higher proportion of weak hydrogen bonds due to the activation of embryonic development. The embryogenic activity could be assessed through the water hydrogen bond network, which is affected by newly generated proteins with different secondary structures.

In ovo exposure to brominated flame retardants Part I: Assessment of effects of TBBPA-BDBPE on survival, morphometric and physiological endpoints in zebra finches.

Tetrabromobisphenol A bis(2,3-dibromopropyl) ether (TBBPA-BDBPE) is an additive flame retardant used in polyolefins and polymers. It has been detected in biota, including in avian eggs, yet little is known of its effects. We assessed the pattern of TBBPA-BDBPE concentrations in songbird eggs over the incubation period, and the effects of embryonic exposure to TBBPA-BDBPE in a model songbird species, the zebra finch (Taeniopygia guttata). To assess concentrations during embryo development, eggs were injected on the day they were laid with the vehicle control (safflower oil) or 100 ng TBBPA-BDBPE/g egg, and whole egg contents were collected throughout embryonic development on day 0 (unincubated), 5, 10 and 13. To evaluate effects of embryonic exposure to TBBPA-BDBPE, eggs were injected at Hamburger-Hamilton stage 18 (∼80 h after initiation of incubation) with safflower oil only, 10, 50 or 100 ng TBBPA-BDBPE/g egg (albumin injection volume 1 µl/g). Eggs were monitored for hatching success, and nestlings were monitored for growth and survival. At 15 days post-hatch, tissues were collected to assess physiological effects. TBBPA-BDBPE was incorporated into the egg as the embryo developed, and concentrations started declining in late incubation, suggesting biotransformation by the embryo. There were no effects on hatching success, nestling survival, growth, organ somatic indices, or thyroid hormone homeostasis; however, there was evidence that body condition declined in a dose-dependent manner towards the end of the rapid nestling growth phase. This decreased body condition could be a delayed effect of early developmental exposure, or it may be the result of increased exposure to biotransformation products of TBBPA-BDBPE produced over the nestling period, which are predicted to be more bioaccumulative and toxic than the parent compound.

In ovo exposure to brominated flame retardants Part II: Assessment of effects of TBBPA-BDBPE and BTBPE on hatching success, morphometric and physiological endpoints in American kestrels.

Tetrabromobisphenol A bis(2,3-dibromopropyl ether) (TBBPA-BDBPE) and 1,2-bis(2,4,6-tribromophenoxy)ethane (BTPBE) are both brominated flame retardants (BFRs) that have been detected in birds; however, their potential biological effects are largely unknown. We assessed the effects of embryonic exposure to TBBPA-BDBPE and BTBPE in a model avian predator, the American kestrel (Falco sparverius). Fertile eggs from a captive population of kestrels were injected on embryonic day 5 (ED5) with a vehicle control or one of three doses within the range of concentrations that have been detected in biota (nominal concentrations of 0, 10, 50 or 100 ng/g egg; measured concentrations 0, 3.0, 13.7 or 33.5 ng TBBPA-BDBPE/g egg and 0, 5.3, 26.8 or 58.1 ng BTBPE/g egg). Eggs were artificially incubated until hatching (ED28), at which point blood and tissues were collected to measure morphological and physiological endpoints, including organ somatic indices, circulating and glandular thyroid hormone concentrations, thyroid gland histology, hepatic deiodinase activity, and markers of oxidative stress. Neither compound had any effects on embryo survival through 90% of the incubation period or on hatching success, body mass, organ size, or oxidative stress of hatchlings. There was evidence of sex-specific effects in the thyroid system responses to the BTBPE exposures, with type 2 deiodinase (D2) activity decreasing at higher doses in female, but not in male hatchlings, suggesting that females may be more sensitive to BTBPE. However, there were no effects of TBBPA-BDBPE on the thyroid system in kestrels. For the BTPBE study, a subset of high-dose eggs was collected throughout the incubation period to measure changes in BTBPE concentrations. There was no decrease in BTBPE over the incubation period, suggesting that BTBPE is slowly metabolized by kestrel embryos throughout their ∼28-d development. These two compounds, therefore, do not appear to be particularly toxic to embryos of the American kestrel.

A pulsed-dose study evaluating chronic toxicity of chlorothalonil to fish: A case study for environmental risk assessment.

Chlorothalonil is a fungicide which is highly toxic to aquatic organisms. However, in natural aquatic environments, it is very rapidly degraded, with a half-life typically in hours, reducing exposure of aquatic organisms and the potential for effects. In standard regulatory studies looking at the chronic toxicity of chlorothalonil to fathead minnow, the most sensitive endpoint was fecundity. A standard fish full-life cycle study, where chlorothalonil concentrations were maintained constant throughout, resulted in a no-observed-effect concentration (NOEC) of 1.4 µg/L. Comparing peak modeled exposure concentrations to this NOEC can result in the chronic risk to fish being considered unacceptable. The present study investigated the effect on fecundity in fathead minnow using a fish short-term reproduction assay. Five different exposure profiles were employed with time-varying concentrations based on realistic worst-case modeled environmental exposure profiles, multiplied by an assessment factor of 10, which resulted in maximum measured concentrations up to 15.5 µg/L. There were no effects on fecundity from any of the exposure profiles tested. Therefore, based on these more realistic exposure profiles, the chronic risk to fish could be considered acceptable if these exposures were deemed to be representative of the worst case. Environ Toxicol Chem 2019;38:1549-1559. © 2019 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

Allocation of glycerolipids and glycerophospholipids from adults to eggs in Daphnia magna: Perturbations by compounds that enhance lipid droplet accumulation.

Analysis of the disruptive effects of chemicals on lipids in invertebrates is limited by our poor knowledge of the lipid metabolic pathways and the complete lipidome. Recent studies shown that juvenoids and bisphenol A disrupted the dynamics of lipid droplets in the crustacean Daphnia magna. This study used ultra-high performance liquid chromatography/time-of-flight mass spectrometry (UHPLC/TOFMS) to study how juvenoids (pyriproxyfen and methyl farnesoate) and bisphenol A disrupt the dynamics of glycerophospholipids and glycerolipids in Daphnia adults and their allocation to eggs. Lipidomic analysis identified 234 individual lipids corresponding to three classes of glycerolipids, seven of glycerophospholipids, and one of sphingolipids, of which 194 changed according to the chemical treatments and time. Adult females in the control and bisphenol A treatment groups had low levels of triacylglycerols but high levels of glycerophospholipids, whereas those in the juvenoid treatment groups had high levels of triacylglycerols and low levels of glycerophospholipids. The opposite trend was observed for the lipid contents in the eggs produced. Because the juvenoids reduced reproduction dramatically, the females allocated less triacylglycerols to their eggs than the controls did. Interestingly, females exposed to bisphenol A allocated less triacylglycerols to their eggs despite producing a similar number of eggs as that of the controls. Thin-layer chromatography analyses confirmed the UHPLC/TOFMS results and allowed qualitative determination of cholesterol, which was also accumulated in females exposed to the juvenoids.

Same medicine, different reasons: Comparing women's bodily experiences of producing eggs for pregnancy or for profit.

Women doing in vitro fertilization (IVF) to have a child describe it as painful and emotionally draining. Egg donors undergo the same medical regimen for a different reason - to produce eggs for another woman in exchange for thousands of dollars - and describe it as quick and relatively painless. Medical researchers typically compare bodily responses by variables such as gender, age, and health status. We use the case of "egg production" to propose a new factor that may be an important source of variation in bodily experience: an individual's reason for undergoing the medical intervention in the first place. Using cluster analysis to analyze an original survey of 50 IVF patients and 62 egg donors from the United States, we find two distinct kinds of bodily experiences - "less intense" and "more intense" - and the intensity of one's experience is associated with one's reason for producing eggs: either to become pregnant or to donate them for money.

Matching Dietary Amino Acid Balance to the In Silico-Translated Exome Optimizes Growth and Reproduction without Cost to Lifespan.

Balancing the quantity and quality of dietary protein relative to other nutrients is a key determinant of evolutionary fitness. A theoretical framework for defining a balanced diet would both reduce the enormous workload to optimize diets empirically and represent a breakthrough toward tailoring diets to the needs of consumers. Here, we report a simple and powerful in silico technique that uses the genome information of an organism to define its dietary amino acid requirements. We show for the fruit fly Drosophila melanogaster that such "exome-matched" diets are more satiating, enhance growth, and increase reproduction relative to non-matched diets. Thus, early life fitness traits can be enhanced at low levels of dietary amino acids that do not impose a cost to lifespan. Exome matching also enhanced mouse growth, indicating that it can be applied to other organisms whose genome sequence is known.

First assessment of trace metal concentration in mangrove crab eggs and other tissues, SE Brazil.

The mangrove crab Ucides cordatus is widespread in the Brazilian coast, which has an important role in nutrient cycling. This species reproduces in summer and females carry eggs about a month, when they maintain contact with water and sediments. It remains unclear if trace metals can be absorbed or adsorbed by the eggs during development. The present study aims to investigate, for the first time, trace metal concentrations in ovigerous female tissues and eggs of U. cordatus in two areas with different metal pollution levels in the Southeastern Brazil. Samples were collected in two different mangroves, Guanabara Bay (GB) highly polluted environment and Paraíba do Sul River (PSR). In both populations, we observed significant increase of V, Cr, and Mn concentrations along eggs maturation. The higher metals averages were found in PSR population. This trend was reported since the 1990s and lower concentrations in GB marine organisms were attributed to reducing conditions, high organic load, and the presence of sulfide ions. These conditions restrict the bioavailability of metals in the bay, with exception of Mn. No significant differences were observed in gills and muscles. In both populations of the present study, V, Zn, As, and Pb were higher in eggs of initial stage, whereas Mn, Ni, Cu, and Cd were higher in hepatopancreas. Beside this, V, Cr, Mn, As, and Pb showed an increase concerning egg development. Thus, V, As, and Pb in eggs come from two sources previous discussed: females and environment. Zinc came mainly from females due to essential function. Those new information should be considered as one of the mechanisms of trace metal transfer to the trophic chain, between benthonic and pelagic environment.

Clupeiformes' Egg Envelope Proteins characterization: The case of Engraulis encrasicolus as a proxy for stock assessment through a novel molecular tool.

Zona radiata proteins are essential for ensuring bactericidal resistance, oocyte nutrients uptake and functional buoyancy, sperm binding and guidance to the micropyle, and protection to the growing oocyte or embryo from the physical environment. Such glycoproteins have been characterized in terms of molecular structure, protein composition and phylogenetics in several chordate models. Nevertheless, research on teleost has not been extensive. In Clupeiformes, one of the most biologically relevant and commercially important order which accounts for over 400 species and totally contributes to more than a quarter of the world fish catch, Egg Envelope Protein (EEP) information exist only for the Clupea pallasii and Engraulis japonicus species. The European anchovy, Engraulis encrasicolus, the target of a well-consolidated fishery in the Mediterranean Sea, has been ignored until now and the interest on the Otocephala superorder has been fragmentally limited to some Cypriniformes and Gonorynchiformes, as well. The aim of the present study was to fill the ZP protein-wise gap of knowledge afflicting the understanding of the European anchovy's reproductive process and to expand the background on Clupeiformes. We cloned the five Engraulis encrasicolus' zp genes and deduced their products, determined their tissue distribution, quantified their mRNA expression throughout the reproductive cycle and provided an insight into their evolution through phylogenetic tools. Furthermore, we proposed a multivariate statistics-based method to objectively infer and/or confirm the classification of Engraulis encrasicolus' sexual maturity stages by analyzing data of zp mRNAs' relative abundance.

Assessment of bagging GBLUP for whole-genome prediction of broiler chicken traits.

Bootstrap aggregation (bagging) is a resampling method known to produce more accurate predictions when predictors are unstable or when the number of markers is much larger than sample size, because of variance reduction capabilities. The purpose of this study was to compare genomic best linear unbiased prediction (GBLUP) with bootstrap aggregated sampling GBLUP (Bagged GBLUP, or BGBLUP) in terms of prediction accuracy. We used a 600 K Affymetrix platform with 1351 birds genotyped and phenotyped for three traits in broiler chickens; body weight, ultrasound measurement of breast muscle and hen house egg production. The predictive performance of GBLUP versus BGBLUP was evaluated in different scenarios consisting of including or excluding the TOP 20 markers from a standard genome-wide association study (GWAS) as fixed effects in the GBLUP model, and varying training sample sizes and allelic frequency bins. Predictive performance was assessed via five replications of a threefold cross-validation using the correlation between observed and predicted values, and prediction mean-squared error. GBLUP overfitted the training set data, and BGBLUP delivered a better predictive ability in testing sets. Treating the TOP 20 markers from the GWAS into the model as fixed effects improved prediction accuracy and added advantages to BGBLUP over GBLUP. The performance of GBLUP and BGBLUP at different allele frequency bins and training sample sizes was similar. In general, results of this study confirm that BGBLUP can be valuable for enhancing genome-enabled prediction of complex traits.

Temperature-independent energy expenditure in early development of the African clawed frog Xenopus laevis.

The thermal dissipation of activated eggs and embryos undergoing development from cleavage to the tailbud stage of the African clawed frog Xenopus laevis was measured as a function of incubation time at temperatures ranging from T = 288.2 K to 295.2 K, using a high-precision isothermal calorimeter. A23187-mediated activation of mature eggs induced stable periodic thermal oscillations lasting for 8-34 h. The frequency agreed well with the cell cycle frequency of initial cleavages at the identical temperature. In the developing embryo, energy metabolism switches from embryonic to adult features during gastrulation. The thermal dissipation after gastrulation fit well with a single modified Avrami equation, which has been used for modeling crystal-growth. Both the oscillation frequency of the activated egg and the growth rate of the embryo strongly depend on temperature with the same apparent activation energy of approximately 87 kJ mole(-1). This result suggests that early development proceeds as a single biological time, attributable to a single metabolic rate. A temperature-independent growth curve was derived by scaling the thermogram to the biological time, indicating that the amount of energy expenditure during each developmental stage is constant over the optimal temperature range.

Isolation and assessment of signaling proteins from synchronized cultures during egg activation and through the egg-to-embryo transition in sea urchins.

Sea urchins are an excellent model system for investigating fertilization mechanisms and fundamental cell biological phenomenon such as release from quiescence, cell division, secretion, and basic signal transduction. The ease of gamete collection, fertilization, and culture is complemented by exquisite developmental synchronicity and the ability to carry out both large-scale biochemical studies and single-cell experiments. In particular, fertilization in echinoderms serves as a paradigm for a digital signaling event-a one-time only switch that launches the egg into the developmental pathway. Sperm-induced egg activation is dependent on the release of calcium from internal stores and subsequent effects on a myriad of cellular events such as exocytosis, cytoskeletal remodeling, and cell cycle reentry. Here we describe methods to investigate individual signaling proteins as well as global proteomic and phosphoproteomic changes involved in the initial steps of egg activation through the egg-to-embryo transition.

Immunological and reproductive health assessment in herring gulls and black-crowned night herons in the Hudson-Raritan Estuary.

Previous studies have shown inexplicable declines in breeding waterbirds within western New York/New Jersey Harbor between 1996 and 2002 and elevated polychlorinated dibenzo-p-dioxins and polychlorinated biphenyls (PCBs) in double-crested cormorant (Phalacrocorax auritus) eggs. The present study assessed associations between immune function, prefledgling survival, and selected organochlorine compounds and metals in herring gulls (Larus argentatus) and black-crowned night herons (Nycticorax nycticorax) in lower New York Harbor during 2003. In pipping gull embryos, lymphoid cells were counted in the thymus and bursa of Fabricius (sites of T and B lymphocyte maturation, respectively). The phytohemagglutinin (PHA) skin response assessed T cell function in gull and heron chicks. Lymphocyte proliferation was measured in vitro in adult and prefledgling gulls. Reference data came from the Great Lakes and Bay of Fundy. Survival of prefledgling gulls was poor, with only 0.68 and 0.5 chicks per nest surviving to three and four weeks after hatch, respectively. Developing lymphoid cells were reduced 51% in the thymus and 42% in the bursa of gull embryos from New York Harbor. In vitro lymphocyte assays demonstrated reduced spontaneous proliferation, reduced T cell mitogen-induced proliferation, and increased B cell mitogen-induced proliferation in gull chicks from New York Harbor. The PHA skin response was suppressed 70 to 80% in gull and heron chicks. Strong negative correlations (r = -0.95 to -0.98) between the PHA response and dioxins and PCBs in gull livers was strong evidence suggesting that these chemicals contribute significantly to immunosuppression in New York Harbor waterbirds.

Polychlorinated biphenyls and organochlorine pesticides in local waterbird eggs from Hong Kong: risk assessment to local waterbirds.

The contamination status of the marine environment in Hong Kong was studied by measuring concentrations of organochlorine (OC) pollutants (i.e., hexachlorobenzene, aldrin, dieldrin, endrin, mirex, total heptachlor, total chlordane, total DDTs, total PCBs, and total toxaphenes) in the eggs of selected waterbird species from different locations around the city: Little Egret (Egretta garzetta) and Chinese Pond Heron (Ardeola bacchus) from Mai Po Village, Great Egret (Ardea alba) and Black-crowned Night Heron (Nycticorax nycticorax) from A Chau, and Chinese Pond Heron (A. bacchus) from Ho Sheung Heung. The mean concentrations of total PCBs and total DDTs ranged from 191-11,100 ng g(-1) lipid and 453-49,000 ng g(-1) lipid, respectively. Recent exposure of waterbirds to technical chlordane was found in Hong Kong. The risk characterization demonstrated potential risks to birds associated with exposure to DDE, which was found to cause a reduction in survival of young in Hong Kong Ardeids based on the endpoint in the risk assessment.

Broodstock diet effect on sea urchin Paracentrotus lividus (Lamarck, 1816) endotrophic larvae development: potential for their year-round use in environmental toxicology assessment.

The effect of captive broodstock diet on fertilization and endotrophic larvae development of the sea urchin Paracentrotus lividus was assessed. Maize grain and five inert pelleted diets were tested, during a three-month experimental period. Maize flour, wheat flour, soybean flour, maize/wheat flour (MWF) and maize/soybean flour mixes were used as vegetal sources for inert feed. Gonad index, percent egg fertilization and larvae malformation occurrence were compared with the results obtained from wild sea urchins (W). Whole egg total amino acid composition was concomitantly analyzed as a tool to explain eventual endotrophic larvae malformations caused by lack of specific nutrients. For all treatment groups (wild and captive), percent egg fertilization values above 96% were always observed, fulfilling the requisites (70-90%) necessary to conduct environmental monitoring bioassays, according to USEPA (2002). Similar values for normal percent larval development were only obtained from P. lividus broodstock subjected to an inert feeding diet based on a maize/wheat flour mix (85.0±1.45%), in comparison to wild P. lividus (82.5±1.75%). Likewise, no statistical differences on resultant whole egg total amino acid composition were observed between P. lividus fed MWF and wild treatments. Moreover, statistical differences between MWF and all the other captive feeding treatments were found for six out of the seventeen amino acids analyzed. This study demonstrates the possibility to obtain high values for P. lividus endotrophic larvae percent normal development based on broodstock held in captivity as long as an appropriate inert diet is provided.

Tissue-based risk assessment of great blue heron (Ardea herodias) exposed to PCDD/DF in the Tittabawassee River floodplain, Michigan, USA.

Concentrations of dioxin-like compounds, primarily polychlorinated dibenzofurans (PCDFs), in soils and sediments of the Tittabawassee River (TR) and associated floodplains downstream of Midland, Michigan, USA, were greater than upstream sites and prompted a site-specific risk assessment of great blue herons (GBH). Tissue exposure of PCDF and polychlorinated dibenzo-p-dioxins (PCDDs) was assessed in multiple GBH tissue types, including blood plasma of adults and eggs, as well as blood plasma, adipose, liver, and muscle of nestlings. Adult GBH exposure was associated with foraging area and age class, with concentrations of PCDD/DF being greater in blood plasma of adult GBH foraging in the TR compared with those foraging in upstream reference areas and in older birds as compared with their younger cohorts. Concentrations of PCDD/DFs and dioxin-like polychlorinated biphenyls (PCBs) in eggs and nestling tissues of GBH collected from rookeries within the TR floodplain were generally similar among rookeries. Mean concentrations of PCDD/DFs in eggs of GBH ranged from 45 to 67 ng/kg, wet weight for the rookeries studied, with a maximum concentration of 210 ng/kg, wet weight observed. Adipose consistently had the greatest concentration of PCDD/DFs of all tissues collected from nestlings of GBH, ranging from 98 to 430 ng/kg, wet weight. Potential for adverse population-level effects from site-specific contaminant exposures were evaluated by comparison with selected toxicity reference values (TRVs). Minimal risk of adverse population-level effects were predicted when exposures measured in tissues of GBH collected from rookeries within the TR were compared with appropriate TRVs. This prediction is consistent with site-specific measures of population condition, which included clutch size and number of nestlings per successful nest.

Influence of avian reproduction ecotoxicological endpoints in the assessment of plant protection products.

The aim of this paper is to examine the statistical relevance of bird species on the endpoints of avian long-term toxicity studies (eggs laid, eggs set, eggs hatching, embryo survivor, 14-day old survivors and eggshell thickness). Data from 561 animals of three different species (Colinus virginianus, Anas platyrynchos and Coturnix coturnix japonica) tested with five different pesticides were analyzed in this study. The substances considered were: Thiamethoxam (EZ-3-(2-chloro-1,3-thiazol-5-ylmethyl)-5-methyl-1,3,5-oxadiazinan-4-ylidene(nitro)amine), Thiacloprid ((Z)-3-(6-chloro-3-pyridylmethyl)-1,3-thiazolidin-2-ylidenecyanamide), Acetamiprid ((E)-N(1)-[(6-chloro-3-pyridyl)methyl]-N(2)-cyano-N(1)-methylacetamidine), Phosmet (O,O-dimethyl S-phthalimidomethyl phosphorodithioate) and Dicofol (2,2,2-trichloro-1,1-bis(4-chlorophenyl)ethanol). Several general lineal mixed models were conducted to evaluate the factors affecting variables used in long-term reproductive toxicity tests. Test significance was p < 0.01 in all models tested. Model R(2) value was high (0.80) for all variables except for eggs laid (R(2) = 0.42) for the three species studied. Tukey studentized range test showed significant differences among species and pesticides. For pre-hatching period the differences were significant for eggs laid and eggs set among species. C. japonica showed statistical differences for egg hatching. With respect to embryo survivor and 14 days old survival, significant different were found for C.virginianus and A. platyrynchos, respectively. These results indicate that the selected species have an influence in the endpoints to be used for risk assessment.

Assessment of sperm chemokinesis with exposure to jelly coats of sea urchin eggs and resact: a microfluidic experiment and numerical study.

Specific peptides contained within the extracellular layer, or jelly coat, of a sea urchin egg have been hypothesized to play an important role in fertilization, though separate accounting of the effects of chemoattraction, chemokinesis, sperm agglomeration and the other possible roles of the jelly coat have not been reported. In the present study, we used a microfluidic device that allowed determination of the differences in the diffusion coefficients of sperm of the purple sea urchin Arbacia punctulata subjected to two chemoattractants, namely the jelly coat and resact. Our objectives were twofold: (1) to experimentally determine and compare the diffusion coefficients of Arbacia punctulata spermatozoa in seawater, jelly coat solution and resact solution; and (2) to determine the effect of sea urchin sperm diffusion coefficient and egg size on the sperm-egg collision frequency using stochastic simulations. Numerical values of the diffusion coefficients obtained by diffusing the spermatozoa in seawater, resact solution and jelly coat solution were used to quantify the chemotactic effect. This allowed direct incorporation of known enlargements of the egg, and altered sperm diffusion coefficients in the presence of chemoattractant, in the stochastic simulations. Simulation results showed that increase in diffusion coefficient values and egg diameter values increased the collision frequency. From the simulation results, we concluded that type of sperm, egg diameter and diffusion coefficient are significant factors in egg fertilization. Increasing the motility of sperm appears to be the prominent role of the jelly coat.